Histatin for corneal wound healing and ocular surface disease

ABSTRACT

Histatins may be used for corneal wound healing and as a treatment for ocular surface disease in humans and other animals. For example, histatins could be included in eye drops, eye gels, ointment, glue, or embedded in (polymer) contact lenses.

REFERENCE TO RELATED APPLICATIONS

This is a continuation application of co-pending application Ser. No.15/180,476, filed Jun. 13, 2016, entitled “HISTATIN FOR CORNEAL WOUNDHEALING AND OCULAR SURFACE DISEASE” which claims priority fromapplication Ser. No. 13/788,803, filed Mar. 7, 2013, entitled “HISTATINFOR CORNEAL WOUND HEALING AND OCULAR SURFACE DISEASE”, which is nowabandoned and which claims one or more inventions which were disclosedin Provisional Application No. 61/648,845, filed May 18, 2012, entitled“HISTATIN FOR CORNEAL WOUND HEALING AND OCULAR SURFACE DISEASE”. Thebenefit under 35 USC § 119(e) of the United States provisionalapplication is hereby claimed, and the aforementioned applications arehereby incorporated herein by reference.

BACKGROUND OF THE INVENTION Field of the Invention

The invention pertains to the field of wound and disease healing. Moreparticularly, the invention pertains to corneal wound healing andtreating ocular surface disease using histatins.

Description of Related Art

Histatins have been shown in in vitro studies to be wound healing agentsfrom saliva. More specifically, WO 2009/087117 (and its US equivalent USPatent Publication 2011/0178010), herein incorporated by reference,identified peptides of histatin, which had wound healing properties invitro.

Histatin 1 (Hst-1) and Histatin 2 (Hst-2) have been identified as majorwound-closing factors in human saliva (“Discovery of the Wound HealingCapacity of Salivary Histatins”, thesis of Menno Johannes Oudhoff,Academic Centre for Dentistry Amsterdam (ACTA), VU University Amsterdamand University of Amsterdam, The Netherlands, 2010, herein incorporatedby reference). These studies were all done in vitro and can not betranslated to a finding for therapeutic or clinical use, especiallysince wound and disease healing are complex processes that need to behighly regulated in order to function properly.

SUMMARY OF THE INVENTION

Histatins may be used for corneal wound healing and as a treatment forocular surface disease in humans and other animals. For example,histatins could be included in eye drops, eye gels, ointment, glue, orembedded in (polymer) contact lenses.

In one preferred embodiment, a method of treating corneal woundsincludes the step of administering a therapeutic amount of at least apeptide fragment of a histatin at a site of a corneal wound. Thehistatin is preferably administered using eye drops, gels, ointmentsincluding histatin, tissue glue, or by incorporating histatin into acontact lens worn by a patient. In a preferred embodiment, thetherapeutic amount of histatin accelerates wound healing compared tocorneal wounds not treated with histatin. The peptide fragment of thehistatin preferably includes a sequence selected from the groupconsisting of: SEQ ID NO: 1; SEQ ID NO: 2; SEQ ID NO: 3; SEQ ID NO: 4;SEQ ID NO: 5; SEQ ID NO: 6; SEQ ID NO: 7; SEQ ID NO: 8; SEQ ID NO: 9;SEQ ID NO: 10; SEQ ID NO: 11; SEQ ID NO: 12; SEQ ID NO: 13; SEQ ID NO:14; SEQ ID NO: 15; SEQ ID NO: 16; SEQ ID NO: 17; SEQ ID NO: 18; SEQ IDNO: 19; SEQ ID NO: 20; SEQ ID NO: 21; SEQ ID NO: 22; SEQ ID NO: 23; SEQID NO: 24; SEQ ID NO: 25; SEQ ID NO: 26; SEQ ID NO: 27; SEQ ID NO: 28;SEQ ID NO: 29; SEQ ID NO: 30; SEQ ID NO: 31; SEQ ID NO: 32; SEQ ID NO:33; and any combination of SEQ ID NO: 1 through SEQ ID NO: 33.

In some preferred embodiments, the histatin includes a) at least apeptide fragment of histatin 5 and b) at least a peptide fragment ofhistatin 1, at least a peptide fragment of histatin 2 or a combinationof at least a peptide fragment of histatin 1 and at least a peptidefragment of histatin 2. In other preferred embodiments, the histatin isat least a peptide fragment of histatin 1, at least a peptide fragmentof histatin 2, at least a peptide fragment of histatin 5, or anycombination of a peptide fragment of histatin 1, a peptide fragment ofhistatin 2 and a peptide fragment of histatin 5.

In other preferred embodiments, the histatin is histatin 1 (SEQ ID NO:4), histatin 2 (SEQ ID NO: 5), histatin 5 (SEQ ID NO: 30), or anycombination of histatin 1, histatin 2, and histatin 5. In otherpreferred embodiments, the histatin includes a) histatin 5 (SEQ ID NO:30) and b) histatin 1 (SEQ ID NO: 4), histatin 2 (SEQ ID NO: 5), or acombination of histatin 1 and histatin 2.

In another preferred embodiment, a therapeutic amount of at least apeptide fragment of a histatin is administered to an ocular surface totreat ocular surface disease. The histatin is preferably administeredusing eye drops, gels, ointments including histatin, tissue glue, or byincorporating histatin into a contact lens worn by a patient. In apreferred embodiment, the therapeutic amount of histatin accelerateshealing of ocular surface disease compared to ocular surface disease nottreated with histatin. The peptide fragment of the histatin preferablyincludes a sequence selected from the group consisting of: SEQ ID NO: 1;SEQ ID NO: 2; SEQ ID NO: 3; SEQ ID NO: 4; SEQ ID NO: 5; SEQ ID NO: 6;SEQ ID NO: 7; SEQ ID NO: 8; SEQ ID NO: 9; SEQ ID NO: 10; SEQ ID NO: 11;SEQ ID NO: 12; SEQ ID NO: 13; SEQ ID NO: 14; SEQ ID NO: 15; SEQ ID NO:16; SEQ ID NO: 17; SEQ ID NO: 18; SEQ ID NO: 19; SEQ ID NO: 20; SEQ IDNO: 21; SEQ ID NO: 22; SEQ ID NO: 23; SEQ ID NO: 24; SEQ ID NO: 25; SEQID NO: 26; SEQ ID NO: 27; SEQ ID NO: 28; SEQ ID NO: 29; SEQ ID NO: 30;SEQ ID NO: 31; SEQ ID NO: 32; SEQ ID NO: 33; and any combination of SEQID NO: 1 through SEQ ID NO: 33.

In some preferred embodiments, the histatin includes a) at least apeptide fragment of histatin 5 and b) at least a peptide fragment ofhistatin 1, at least a peptide fragment of histatin 2 or a combinationof at least a peptide fragment of histatin 1. In other preferredembodiments, the histatin is at least a peptide fragment of histatin 1,at least a peptide fragment of histatin 2, at least a peptide fragmentof histatin 5 or any combination of a peptide fragment of histatin 1, apeptide fragment of histatin 2 and a peptide fragment of histatin 5.

In other preferred embodiments, the histatin is histatin 1 (SEQ ID NO:4), histatin 2 (SEQ ID NO: 5), histatin 5 (SEQ ID NO: 30), or anycombination of histatin 1, histatin 2, and histatin 5. In otherpreferred embodiments the histatin includes a) histatin 5 (SEQ ID NO:30) and b) histatin 1 (SEQ ID NO: 4), histatin 2 (SEQ ID NO: 5), or acombination of histatin 1 and histatin 2.

DETAILED DESCRIPTION OF THE INVENTION

Histatins are naturally occurring oral peptides produced by humans andnon-human primates that demonstrate direct anti-infective activity,potent anti-inflammatory properties, and stimulate epithelial woundhealing in several tissue and organ culture systems. A research facilityhas developed a technique to isolate this natural substance, making it apotential topical treatment for wounds.

In a preferred embodiment, a peptide including at least one amino acidsequence of at least eight amino acids adjacently present in Histatin 1,2, 3, and/or 5 is used to treat a corneal wound or ocular surfacedisease.

In one preferred embodiment, a method of treating corneal woundsincludes the step of administering a therapeutic amount of at least aportion of a histatin peptide at a site of a corneal wound. In anotherpreferred embodiment, a method of treating ocular surface diseaseincludes the step of administering a therapeutic amount of at least aportion of a histatin peptide to an ocular surface. The ocular surfacediseases may include, but are not limited to, dry eyes, cornealulcerations and erosions, inflammatory and infectious keratitis andconjunctivitis, surgical interventions, and trauma.

The histatin is preferably administered using eye drops, gels, ointmentsincluding histatin, tissue glue, or by incorporating histatin into acontact lens worn by a patient. In a preferred embodiment, thetherapeutic amount of histatin accelerates wound or ocular surfacedisease healing compared to corneal wounds or ocular surface diseasesnot treated with histatin.

In some preferred embodiments, the histatin concentration is betweenapproximately 0.1 μg/ml and approximately 1000 mg/ml. In other preferredembodiments, the histatin concentration is between approximately 0.1μg/ml and 10 μg/ml. In some preferred embodiments, the histatinconcentration is greater than or equal to approximately 1 μM.

The administering step may be repeated multiple times per day and/or fora plurality of days. In one preferred embodiment, this step is repeatedat least one time a day for a plurality of days. In another preferredembodiment, the step is repeated chronically at least one time a day. Insome preferred embodiments, the step is repeated up to hourly for aplurality of days. In another preferred embodiment, the step is repeatedat least two times a day for a plurality of days. In yet anotherpreferred embodiment, the step is repeated at least three times a dayfor a plurality of days, for example for seven days. In anotherpreferred embodiment, the step is repeated four times a day for fivedays.

In one preferred embodiment, the histatin is a peptide including 8 to 44amino acids. In some preferred embodiments, the peptide is a L-peptide.In other preferred embodiments, the peptide is a cyclic peptide.

In some preferred embodiments, the amino acid sequence of the histatinpeptide is one or more of SEQ ID NOS: 1 through 33, or any combinationsof these sequences. In alternative embodiments, one or more of the aminoacid sequences have a substitution, deletion and/or insertion of up to 3amino acids. In other alternative embodiments, one or more of the aminoacid sequences have a substitution, a deletion and/or an insertion oftwo or less amino acids. In other alternative embodiments, one or moreof the amino acid sequences have a substitution, a deletion, and/or aninsertion in one amino acid.

The SEQ ID NO: 4 peptide is also known as Histatin 1 (Hst-1). Note thatthe first serine in this amino acid sequence may be a phosphoserine. TheSEQ ID NO: 5 peptide is also known as Histatin 2 (Hst-2, also equivalentto amino acids 12-38 of Hst-1). The SEQ ID NO: 6 peptide is also knownas Histatin 3 (Hst-3). The SEQ ID NO: 30 peptide is also known asHistatin 5 (Hst-5). Parts and fragments of each of these amino acidsequences may be used, alone or in combination, including but notlimited to SEQ ID NOS: 1-3, 7-29 (for Histatin 1, Histatin 2 andHistatin 3) and SEQ ID NO: 31 (for Histatin 5) to facilitate woundclosure in the embodiments described herein. While the L stereoisomer ofthe amino acids is preferred for the amino acid sequences describedherein, D stereoisomers may alternatively be used. Alternatively, aminoacid sequences that include these histatins and other amino acids, forexample SEQ ID NO: 33, which is a sortase cyclized histatin (includingall of Histatin 1), may be used in the embodiments described herein. Anyhistatin sequences could be cyclized and used in the embodimentsdescribed herein.

Some preferred embodiments use amino acid sequences from Hst-1 and/orHst-2 in combination with amino acid sequences from Hst-5 to treatcorneal wounds or ocular surface disease. In these embodiments, one ormore amino acid sequences from Hst-1 and/or Hst-2 are chosen, and one ormore amino acid sequences from Hst-5 are chosen. In some embodiments,the full length Histatin 1 (SEQ ID NO: 4), full length Histatin 2 (SEQID NO: 5), and/or the full length Histatin 5 (SEQ ID NO: 30) could beused. In other embodiments, portions of Hst-1, Hst-2, and/or Hst-5 couldbe used. For example, SEQ ID NO: 29, which is equivalent to amino acids20-32 of Histatin 1, may be a preferred amino acid sequence to use forwound closure in some embodiments. In other examples, peptides includingSEQ ID NO: 32, a peptide fragment of Histatin 1 and Histatin 2 thatappears to be a core motif for wound closure, may be used. Otherpreferred sequences from Hst-1 and Hst-2 include, but are not limitedto, SEQ ID NO: 8, SEQ ID NO: 9 and SEQ ID NO: 13. As another example,SEQ ID NO: 31, a fragment of Histatin 5 (Gusman et al., “SalivaryHistatin 5 is an inhibitor of Both Host and Bacterial Enzymes Implicatedin Periodontal Disease”, Infect. Immun 2001, 69(3): 1402, pp. 1402-1408,herein incorporated by reference), may be used, preferably incombination with Histatin 1 or Histatin 2 or fragments thereof. In otherpreferred embodiments, fragments of Hst-1 or Hst-2 are used with fulllength Hst-5 (SEQ ID NO: 30) or full length Hst-1 (SEQ ID NO: 4) orHst-2 (SEQ ID NO: 5) are used with fragments of Hst-5 (for example, SEQID NO: 31). In yet other embodiments, any combination of fragments ofHst-1 and/or Hst-2, full length Hst-1 and/or Hst-2, fragments of Hst-5,or full length Hst-5 may be used. In some preferred embodiments, theconcentration of the Hst-5 peptide used is greater than or equal toapproximately 1 μM.

The amino acids and the peptides described herein may include at leastone functional grouping (for example, an amine and/or carboxylic group)protected with a protective grouping in some embodiments. Since thepeptides are applied to tissue, skin or a wound, a protected form of thepeptide may be preferred to resist degradation. The form of protectionneeds to be biologically compatible and compatible with pharmaceuticaluse. Some examples include, but are not limited to, the acylation or theacetylation of the amino-terminal ends, cyclization or the amidation orthe esterfication of the carboxy-terminal ends. Thus, the peptidesdescribed herein may be used in a protected form.

The peptides described herein may be made by traditional chemicalsynthesis, enzymatic synthesis, or any other method known in the art.

The peptides preferably include at least 8 amino acids. In one preferredembodiment, the peptides include a range of 8 to 44 amino acids, but thepeptides may alternatively include more than 44 amino acids.

In Vivo Studies

Efficacy studies of histatin for corneal wound healing utilize an animalmodel, namely rabbits. Histatins are naturally produced substances thatstimulate healing in several tissue and organ culture systems. Theresults of these studies demonstrate that histatin has a significantdose dependent accelerated healing activity for corneal wounds.

Ocular surface disorders including, but not limited to, dry eyes,conical ulcerations and erosions, inflammatory and infectious keratitisand conjunctivitis, surgical interventions, and trauma all lead todisruptions in the integrity of the conical and conjunctival cellularbarrier that result in increased risk of infection, pain, and reducedvisual acuity. Histatins have a potential use in the treatment of ocularsurface trauma/injury and infectious disease.

The outer layer of the cornea, the corneal epithelium, serves as aphysical barrier against the environment and thus also as a line ofdefense to prevent infectious and/or toxic agents frominfecting/affecting the tissue. When injury occurs to the surface of thecornea, the corneal epithelium spearheads a wound healing process (see,for example, Klyce S D, Crosson C E. “Transport processes across therabbit conical epithelium: a review”. Curr Eye Res. 1985; 4:323-331 andLu L, Wang L, Shell B. “UV-induced signaling pathways associated withcorneal epithelial cell apoptosis”. Invest Ophthalmol Vis Sci. 2003;44:5102-5109, both herein incorporated by reference).

The study disclosed herein evaluates and quantifies the wound healingeffects of histatins on the ocular surface of New Zealand White rabbits.

Brief Methodology:

Epithelial defects are created in the right eye (oculus dexter, OD) of12 New Zealand White rabbits. Due to animal regulations, bilateralwounding is not permitted. After the epithelial defects are made, therabbits are randomized into treatment groups. Two (2) groups are treatedwith different cyclized histatin concentrations: 0.1 μg/ml and 10 μg/mldissolved into an ophthalmic artificial tear preparation and deliveredto the rabbits as an eye drop three times daily. Histatins known in theart, including, but not limited to, amino acid SEQ ID NOS: 1 through 33,which include Hst-1 (SEQ ID NO: 4), Hst-2 (SEQ ID NO: 5), Hst-3 (SEQ IDNO: 6), Hst-5 (SEQ ID NO: 30), and sortase cyclized histatin (SEQ ID NO:33), may be used in these studies or in treatment protocols. One (1)group is treated with an inactive/inert formulation (control). Thiscontrol group should receive the same vehicle identical to the other twogroups but without histatin. An over-the-counter artificial tearpreferably serves as the vehicle. The initial study included four (4)animals/group in three (3) groups, for a total of twelve rabbits.

The groups are to be treated with agent (either histatin or aninactive/inert formulation of artificial tears) three times/day (TID)for 7 days. Each rabbit group is preferably given moxifloxicin treatmentto prevent infection.

The corneal wounds are then evaluated daily for the corneal woundhealing abilities of histatin via fluorescein staining, fluorescent slitlamp biomicrophotography and computerized area determination. Evaluatorsare masked to the therapeutic treatment given to the rabbits. Afterhealing, two (2) animals from each group are euthanized, and the corneascollected for histological processing (H&E staining with subsequentevaluation by veterinary histopathologist). The decision to performhistopathologic analysis after tissue procurement is made only if thereis proven difference in healing between the different treatment groupsand the controls. At study termination (study preferably continues forseven days), the remaining animals are euthanized.

Results

The data from a first study using the methodology above is shown inTable 1. The histatin used in this study was cyclized histatin 1. Thehistatin 1 used was a sortase cyclized histatin with amino acid SEQ IDNO: 33, in which the “C-terminal” T is linked to the “N-terminal” G.Table 2 shows the mm size values (without the standard deviation) as anapproximate percentage of the size at 1 hour post wound for each of thethree groups. Pathological analysis of the rabbit corneas showed notoxicity.

TABLE 1 Hours Post Control 0.1 μg/ml 10 μg/ml Wound (mm²) (mm²) (mm²) 069.3 ± 17.3 112.7 ± 37.8  88.4 ± 31.6 6 72.6 ± 15.2 96.2 ± 13.6 74.0 ±14.7 24 50.6 ± 16.2 77.5 ± 25.0 51.4 ± 9.3  30 46.6 ± 25.8 48.8 ± 8.0 37.7 ± 9.4  48 13.6 ± 20.2 2.7 ± 3.1 0.00 ± 0.07 54 5.2 ± 7.9 0.00 ±0.00 0.00 ± 0.00 72 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00

TABLE 2 Control- 0.1 ug/ml- 10 ug/ml- Percentage Percentage PercentageHours of 1 hour of 1 hour of 1 hour Post post post post Wound wound sizewound size wound size 0 100% 100% 100% 6 105% 85% 84% 24 73% 69% 58% 3067% 43% 43% 48 20% 2% 0% 54 8% 0% 0% 72 0% 0% 0%

The results above show that histatin demonstrates a significant dosedependent accelerated healing activity of corneal wounds. While Table 2does not take into account the standard deviations from Table 1, thepercentages clearly indicate that the wounds treated with 0.1 μg/ml or10 μg/ml histatin healed faster (shrunk more) at each of the time pointswhere data was collected. These results are the first results of theirkind done using in vivo animal studies.

Histatins and peptide portions or peptide fragments of histatins may beused to accelerate corneal wound healing or ocular surface diseasehealing in humans and other animals. In preferred embodiments, histatin1 (Hst-1), histatin 2 (Hst-2), histatin 5 (Hst-5), peptide fragments ofHst1, Hst2, or Hst5, or any combinations thereof may be used. In otherembodiments, histatin 3 (Hst-3) or the D-enantiomer of histatin 2(D-Hst-2), or peptide fragments thereof, may be used. Any combinationsof any of the histatins may be used. In preferred embodiments, histatinconcentrations between 0.1 μg/ml and 1000 mg/ml may be used. Peptideswith amino acid SEQ ID NOS: 1-33, histatins known in the art, thepeptides disclosed in WO 2009/087117 or the peptides disclosed in Dr.Menno Johannes Oudhoff's thesis, “Discovery of the Wound-HealingCapacity of Salivary Histatins”, 2010, department of Oral Biochemistryof the Academic Centre for Dentistry Amsterdam (ACTA), VU UniversityAmsterdam and University of Amsterdam, The Netherlands, hereinincorporated by reference, may be used.

In one preferred embodiment, histatin 1 (Hst-1) or histatin 2 (Hst-2) incombination with histatin 5 (Hst-5), peptide fragments of Hst-1 or Hst-2in combination with peptide fragments of Hst-5, or any combination, areused. Hst-5 inhibits production of Matrix Metalloproteases (MMPs).

The combination of the Hst-1/Hst-2 healing properties with the Hst-5inhibiting MMPs should be very effective. In some preferred embodiments,a concentration of at least approximately 1 μM of Hst-5, or a fragmentof Hst-5, is used.

Histatins could be administered to humans or other animals with acorneal wound or ocular surface disorders. Some methods ofadministration include, but are not limited to, incorporating thehistatin into eye drops, gels or ointments, incorporating the histatininto tissue glue used to transiently seal corneal injuries, or embeddingthe histatin into (polymer) contact lenses.

The histatins may be administered in any combination of daily treatmentsfor any number of days in order to produce therapeutic results. In onepreferred embodiment, the histatin is administered at least once a dayfor a plurality of days. In another preferred embodiment, the histatinis administered at least once a day chronically (for an extended periodof time). In another preferred embodiment, the step may be repeated two,three, four, five times or more, or hourly, for a plurality of days orchronically. In one example, the histatin is repeated three times a dayfor seven days. In another example, histatin is administered four timesa day for five days.

Accordingly, it is to be understood that the embodiments of theinvention herein described are merely illustrative of the application ofthe principles of the invention. Reference herein to details of theillustrated embodiments is not intended to limit the scope of theclaims, which themselves recite those features regarded as essential tothe invention.

What is claimed is:
 1. A method of treating ocular surface disease,comprising the step of continually administering a therapeutic amount ofa composition through incorporation of the composition into a contactlens worn by a patient, the composition comprising a first peptide and asecond peptide to an ocular surface, wherein the first peptide comprisesa first histatin or a fragment of the first histatin and the secondpeptide comprises a second histatin or a fragment of the secondhistatin; wherein the therapeutic amount of the composition is selectedsuch that the composition accelerates healing of ocular surface diseasecompared to healing of ocular surface disease not treated with thecomposition; wherein an amino acid sequence of each of the first peptideand the second peptide is selected from the group consisting of: SEQ IDNO: 1; SEQ ID NO: 2; SEQ ID NO: 3; SEQ ID NO: 4; SEQ ID NO: 5; SEQ IDNO: 6; SEQ ID NO: 7; SEQ ID NO: 8; SEQ ID NO: 9; SEQ ID NO: 10; SEQ IDNO: 11; SEQ ID NO: 12; SEQ ID NO: 13; SEQ ID NO: 14; SEQ ID NO: 15; SEQID NO: 16; SEQ ID NO: 17; SEQ ID NO: 18; SEQ ID NO: 19; SEQ ID NO: 20;SEQ ID NO: 21; SEQ ID NO: 22; SEQ ID NO: 23; SEQ ID NO: 24; SEQ ID NO:25; SEQ ID NO: 26; SEQ ID NO: 27; SEQ ID NO: 28; SEQ ID NO: 29; SEQ IDNO: 30; SEQ ID NO: 31; SEQ ID NO: 32; SEQ ID NO: 33; and any combinationof SEQ ID NO: 1 through SEQ ID NO: 33; wherein the first peptidecomprises histatin 1 or a fragment of histatin 1; and wherein the secondpeptide comprises histatin 5 or a fragment of histatin
 5. 2. The methodof claim 1, wherein the first peptide further comprises histatin 2 or afragment of histatin
 2. 3. The method of claim 1, wherein the methodtreats humans.
 4. The method of claim 1, wherein the concentration ofeach of the first peptide and the second peptide in the composition isbetween approximately 0.1 μg/ml and approximately 1000 mg/ml.
 5. Themethod of claim 4, wherein the concentration of each of the firstpeptide and the second peptide in the composition is betweenapproximately 0.1 μg/ml and 10 μg/ml.
 6. The method of claim 1, furthercomprising repeating the step of administering the therapeutic amount ofthe composition to the ocular surface at least one time per day for aplurality of days.
 7. The method of claim 1, further comprisingchronically repeating the step of administering the therapeutic amountof the composition to the ocular surface at least one time a day.
 8. Themethod of claim 1, wherein the amino acid sequence of the second peptideis selected from the group consisting of SEQ ID NO: 30 and SEQ ID NO:31.
 9. The method of claim 1, wherein the amino acid sequence of thefirst peptide is selected from the group consisting of SEQ ID NO: 4; SEQID NO: 5; SEQ ID NO: 29; and SEQ ID NO:
 33. 10. The method of claim 2,wherein the amino acid sequence of the first peptide is selected fromthe group consisting of SEQ ID NO: 4; SEQ ID NO: 5; SEQ ID NO: 8; SEQ IDNO: 9; SEQ ID NO: 13; SEQ ID NO: 29; and SEQ ID NO:
 33. 11. The methodof claim 1, wherein the ocular surface disease is selected from thegroup consisting of dry eyes, corneal ulcerations and erosions,inflammatory and infectious keratitis and conjunctivitis, surgicalinterventions, and trauma.
 12. The method of claim 1, wherein the firstpeptide and the second peptide are 8 to 44 amino acids in length. 13.The method of claim 1, wherein the first peptide and the second peptideare L-peptides.
 14. The method of claim 1, wherein the first peptide isa cyclic peptide.
 15. The method of claim 14, wherein the first peptidecomprises SEQ ID NO:
 33. 16. The method of claim 1, wherein the firstpeptide comprises SEQ ID NO:
 33. 17. A method of treating ocular surfacedisease, comprising the step of continually administering a therapeuticamount of a composition using eye drops gels, ointments or tissue glue,the composition comprising a first peptide and a second peptide to anocular surface, wherein the first peptide comprises a first histatin ora fragment of the first histatin and the second peptide comprises asecond histatin or a fragment of the second histatin; wherein thetherapeutic amount of the composition is selected such that thecomposition accelerates healing of ocular surface disease compared toocular surface disease not treated with the composition; wherein anamino acid sequence of each of the first peptide and the second peptideis selected from the group consisting of: SEQ ID NO: 1; SEQ ID NO: 2;SEQ ID NO: 3; SEQ ID NO: 4; SEQ ID NO: 5; SEQ ID NO: 6; SEQ ID NO: 7;SEQ ID NO: 8; SEQ ID NO: 9; SEQ ID NO: 10; SEQ ID NO: 11; SEQ ID NO: 12;SEQ ID NO: 13; SEQ ID NO: 14; SEQ ID NO: 15; SEQ ID NO: 16; SEQ ID NO:17; SEQ ID NO: 18; SEQ ID NO: 19; SEQ ID NO: 20; SEQ ID NO: 21; SEQ IDNO: 22; SEQ ID NO: 23; SEQ ID NO: 24; SEQ ID NO: 25; SEQ ID NO: 26; SEQID NO: 27; SEQ ID NO: 28; SEQ ID NO: 29; SEQ ID NO: 30; SEQ ID NO: 31;SEQ ID NO: 32; SEQ ID NO: 33; and any combination of SEQ ID NO: 1through SEQ ID NO: 33; wherein the first peptide comprises histatin 1 ora fragment of histatin 1; and wherein the second peptide compriseshistatin 5 or a fragment of histatin
 5. 18. The method of claim 17,further comprising repeating the step of administering the therapeuticamount of the composition to the ocular surface at least one time perday for a plurality of days.
 19. The method of claim 17, furthercomprising chronically repeating the step of administering thetherapeutic amount of the composition to the ocular surface at least onetime per day.
 20. The method of claim 17, further comprising repeatingthe step of administering the therapeutic amount of the composition tothe ocular surface up to hourly for a plurality of days.